title: Dendritic cells adenovirally-transduced with full-length mesothelin cDNA elicit mesothelin-specific cytotoxicity against pancreatic cancer cell lines in vitro.
abstract: Mesothelin (MSLN) is an attractive candidate as a molecular target for pancreatic cancer immunotherapy. The purpose of this study was to demonstrate that cytotoxic T lymphocytes (CTLs) generated from peripheral blood mononuclear cells (PBMCs) by stimulation with genetically-modified dendritic cells (DCs) expressing MSLN could produce specific anti-tumor immunity against pancreatic cancer cells endogenously expressing MSLN. MSLN-specific CTLs were generated from PBMCs of healthy donors by in vitro stimulation with DCs adenovirally-transduced with the full-length MSLN gene (DC-AxCAMSLN). The cytotoxic activity was tested using a 4-h (51)Cr-release assay. The pancreatic cancer cell lines (PK1, CfPAC1, AsPC1), a lymphoblastoid cell lines (LCL) transduced with the MSLN gene, and LCL pulsed with MSLN-epitope peptides were used as target cells. MSLN-specific CTLs induced by in vitro stimulation with DC-AxCAMSLN killed pancreatic cancer cell lines expressing MSLN in an HLA-restricted fashion. These CTLs also showed cytotoxic activity against autologous LCL pulsed with multiple MSLN-derived epitope peptides. In addition, CD8(+) T cells, as well as CD4(+) T cells, sorted from these CTLs showed significant production of interferon-γ when stimulated with DC-AxCAMSLN. The in vitro stimulation of PBMCs with DCs transduced with the full-length MSLN gene elicited a potent MSLN-specific cytotoxic activity against pancreatic cancer cell lines endogenously expressing MSLN by recognizing multiple MSLN epitopes and activating both CD8(+) T cells and CD4(+) helper T cells. These results therefore suggest the potential of developing future clinical applications of the vaccines using genetically-modified DCs expressing MSLN.
authors.affiliation: Second Department of Surgery, Wakayama Medical University, School of Medicine, Wakayama, Japan.
authors.forename: Motoki
authors.initials: M
authors.lastname: Miyazawa
authors.forename: Makoto
authors.initials: M
authors.lastname: Iwahashi
authors.forename: Toshiyasu
authors.initials: T
authors.lastname: Ojima
authors.forename: Masahiro
authors.initials: M
authors.lastname: Katsuda
authors.forename: Masaki
authors.initials: M
authors.lastname: Nakamura
authors.forename: Mikihito
authors.initials: M
authors.lastname: Nakamori
authors.forename: Kentaro
authors.initials: K
authors.lastname: Ueda
authors.forename: Teiji
authors.initials: T
authors.lastname: Naka
authors.forename: Keiji
authors.initials: K
authors.lastname: Hayata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iida
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamaue
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D018076:DNA, Complementary / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology*; D058851:GPI-Linked Proteins / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D016233:Immunotherapy, Active / Q000379:methods*; D008213:Lymphocyte Activation / Q000235:genetics / Q000276:immunology; D000090204:Mesothelin; D010190:Pancreatic Neoplasms / Q000276:immunology* / Q000628:therapy; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014161:Transduction, Genetic
chemical_list: D019496:Cancer Vaccines; D018076:DNA, Complementary; D058851:GPI-Linked Proteins; C000715527:MSLN protein, human; D000090204:Mesothelin